利用快速瞬时表达试验鉴定蚕蛾幼虫唾液腺中活跃的基因启动子。

IF 3.2 2区 农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Insect Biochemistry and Molecular Biology Pub Date : 2024-07-26 DOI:10.1016/j.ibmb.2024.104163
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引用次数: 0

摘要

组织特异性基因启动子具有控制转基因动物基因表达所需的特异性,因此很受欢迎。在这里,我们描述了一种相对快速的双组分瞬时表达检测方法,该方法用于鉴定活跃于绿孔蝇幼虫唾液腺中的基因启动子。无菌蝇蛆被广泛用于伤口清创。幼虫唾液腺基因启动子可用于制造能分泌促进伤口治疗因子的蝇蛆。向携带四环素转座子(tTA)激活的红色荧光蛋白基因的品系的胚胎注射由组成型或组织特异性基因启动子驱动的 tTA 基因质粒 DNA。孵化出的幼虫经饮食饲养后检测红色荧光。来自 LsCG30371 基因的启动子在幼虫唾液腺中具有活性。随后,携带 LsCG30371-tTA 基因的稳定转基因品系证实了该启动子的组织特异性。这种相对快速的瞬时表达试验可用于确定其他基因启动子的组织特异性。此外,稳定的 LsCG30371-tTA 株系还可用于制造从唾液腺分泌因子以促进伤口愈合的不育蛆。
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Identification of a gene promoter active in Lucilia sericata larval salivary glands using a rapid transient expression assay

Tissue-specific gene promoters are desired as they provide the specificity needed for control of gene expression in transgenic animals. Here we describe a relatively rapid two-component transient expression assay that was used to identify a gene promoter active in the larval salivary glands of the green blow fly, Lucilia sericata. Sterile L. sericata maggots are widely used for wound debridement. A larval salivary gland gene promoter could be used to make maggots that secrete factors for enhanced wound therapy. Embryos from a line that carry a tetracycline transactivator (tTA)-activated red fluorescent protein gene were injected with plasmid DNA with the tTA gene driven by a constitutive or tissue-specific gene promoter. The hatched larvae were reared on diet and then examined for red fluorescence. A promoter from the LsCG30371 gene was active in the larval salivary glands. The tissue-specificity of the promoter was subsequently confirmed with stable transgenic lines that carried the LsCG30371-tTA gene. The relatively rapid transient expression assay could potentially be used to determine the tissue-specificity of other gene promoters. Further, the stable LsCG30371-tTA lines could be used to make sterile maggots that secrete factors from the salivary glands for enhanced wound healing.

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来源期刊
CiteScore
7.40
自引率
5.30%
发文量
105
审稿时长
40 days
期刊介绍: This international journal publishes original contributions and mini-reviews in the fields of insect biochemistry and insect molecular biology. Main areas of interest are neurochemistry, hormone and pheromone biochemistry, enzymes and metabolism, hormone action and gene regulation, gene characterization and structure, pharmacology, immunology and cell and tissue culture. Papers on the biochemistry and molecular biology of other groups of arthropods are published if of general interest to the readership. Technique papers will be considered for publication if they significantly advance the field of insect biochemistry and molecular biology in the opinion of the Editors and Editorial Board.
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